RESUMO
Fascioliasis is a trematodiasis that affects domestic and wild animals as well as humans worldwide. It is a well-recognized disease in livestock, were it produces serious economic losses. Yet in cattle, there is limited information about the burden of liver flukes and its relation to the eggs per gram shed to the environment. There is also lack of knowledge on the effect of parasite load in blood parameters of infected animals, which is important to evaluate the severity and progression of the disease. The objective of this work was to gain insight in these aspects. Cattle from Mendoza province, Argentina, were inspected at a farm and at the abattoir determining the presence or absence of Fasciola hepatica. Each animal was sampled for blood and feces and in the slaughterhouse the livers were inspected. Hematology and blood chemistry parameters were determined, feces were examined for F. hepatica eggs by a quantitative sedimentation technique and livers were thoroughly inspected to determine the number of flukes. Infected cattle presented a mild burden of liver flukes per animal, strongly correlated (r = 0.72) to the number of eggs per gram of feces. The total number of eggs (XÌ=35,100) shed per animal to the environment and the type of livestock management techniques in the region exacerbate the role of cattle as efficient reservoirs of this disease. Statistically significant lower red blood cell, lymphocyte and neutrophil counts were observed in infected compared to uninfected animals. All hepatic parameters tested showed highly statistically significant differences (p < 0.001) as well as proteins by cause of rise of globulins in infected cattle. The correlation between the amount of flukes in the liver and the number of eggs per gram of faces indicates coprology as a reliable and cost-effective method to infer parasite burden. The impact of fascioliasis on blood parameters can be of aid for the veterinary practitioner on the assessment of this disease on cattle.
Assuntos
Doenças dos Bovinos , Fasciola hepatica , Fasciolíase , Fezes , Carga Parasitária , Animais , Fasciolíase/veterinária , Fasciolíase/sangue , Fasciolíase/parasitologia , Bovinos , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/sangue , Doenças dos Bovinos/epidemiologia , Fezes/parasitologia , Fasciola hepatica/isolamento & purificação , Argentina/epidemiologia , Contagem de Ovos de Parasitas , Fígado/parasitologia , Análise Química do Sangue , Doença Crônica , MatadourosRESUMO
E. granulosus is a cestode that causes Cystic Echinococcosis (CE), a zoonotic disease with worldwide presence. The immune response generated by the host against the metacestode induces a permissive Th2 response, as opposed to pro-inflammatory Th1 response. In this view, mixed Th2 and regulatory responses allow parasite survival. Overall, larval Echinococcus infections induce strong regulatory responses. Fasciola hepatica, another common helminth parasite, represents a major infection in cattle. Co-infection with different parasite species in the same host, polyparasitism, is a common occurrence involving E. granulosus and F. hepatica in cattle. 'While it is known that infection with F. hepatica also triggers a polarized Th2/Treg immune response, little is reported regarding effects on the systemic immune response of this example of polyparasitism. F. hepatica also triggers immune responses polarized to the Th2/ Treg spectrum. Serum samples from 107 animals were analyzed, and were divided according to their infection status and Echinococcal cysts fertility. Cytokines were measured utilizing a Milliplex Magnetic Bead Panel to detect IFN-γ, IL-1, IL-2, IL-4, IL-6, IL-10, IL-12 and IL-18. Cattle infected only with F. hepatica had the highest concentration of every cytokine analyzed, with both 4.24 and 3.34-fold increases in IL-10 and IL-4, respectively, compared to control animals, followed by E. granulosus and F. hepatica co-infected animals with two-fold increase in IL-10 and IL-4, compared to control animals, suggesting that E. granulosus co-infection dampens the cattle Th2/Treg immune response against F. hepatica. When considering Echinococcal cyst fertility and systemic cytokine concentrations, fertile cysts had higher IFN-γ, IL-6 and IL-18 concentrations, while infertile cysts had higher IL-10 concentrations. These results show that E. granulosus co-infection lowers Th1 and Th2 cytokine serological concentration when compared to F. hepatica infection alone. E. granulosus infections show no difference in IFN-γ, IL-1, IL-2, IL-6 and IL-18 levels compared with control animals, highlighting the immune evasion mechanisms of this cestode.
Assuntos
Doenças dos Bovinos/epidemiologia , Coinfecção/epidemiologia , Citocinas/sangue , Equinococose/veterinária , Echinococcus granulosus/imunologia , Fasciola hepatica/imunologia , Fasciolíase/veterinária , Animais , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/imunologia , Doenças dos Bovinos/parasitologia , Chile/epidemiologia , Coinfecção/sangue , Coinfecção/imunologia , Coinfecção/parasitologia , Equinococose/sangue , Equinococose/imunologia , Equinococose/parasitologia , Fasciolíase/sangue , Fasciolíase/imunologia , Fasciolíase/parasitologia , Células Th1/imunologia , Células Th2/imunologiaRESUMO
INTRODUCTION: Fascioliasis is a disease caused by the parasite Fasciola hepatica, according to its clinical picture, it can be classified into two phases: acute and chronic, currently the diagnosis is made during the chronic phase, however, early detection would allow to provide an effective treatment and timely. Objetive: This study is carried out with the objective of evaluating the diagnostic validity of serological tests for the timely diagnosis of hepatic fascioliasis. MATERIALS AND METHODS: a systematic search of the literature was carried out in Embase, MedLine, Cochrane Library and LILACS. Study selection was performed by pairs of reviewers, diagnostic test studies were selected using: ELISA, indirect ELISA, rapid ELISA, Fas2 ELISA, Arco2 compared to sedimentation techniques for the diagnosis of hepatic fascioliasis in adults. The studies were evaluated using the QUADAS-2 tool. RESULTS: Seven studies were identified that answer the PICO question, with a total of 1,317 human serum samples, the tests reported high sensitivity and specificity, FhTA, ELISA, number of studies and participants identified, summary estimators with the corresponding confidence intervals / credibility, and the ranking of treatments. DISCUSSION: In general, the specificity of the serological tests was slightly higher than the sensitivity, there is evidence of moderate to low quality that reports their performance, however, the quality of the evidence and the heterogeneity between the studies do not allow determining its usefulness during the acute phase of the disease. More studies are needed in this regard. The studies were clinically heterogeneous between them.
Assuntos
Fasciolíase/sangue , Fasciolíase/diagnóstico , Humanos , Testes SorológicosRESUMO
RESUMEN Introducción: La fascioliasis es una enfermedad causada por el parásito Fasciola hepática, según su cuadro clínico puede clasificarse en dos fases: aguda y crónica, actualmente el diagnóstico se realiza durante la fase crónica, sin embargo, la detección temprana permitiría brindar un tratamiento eficaz y oportuno. Objetivo: Este estudio se realiza con el objetivo de evaluar la validez diagnóstica de las pruebas serológicas para el diagnóstico oportuno de fascioliasis hepática. Materiales y métodos: se realizó una búsqueda sistemática de la literatura en Embase, MedLine, Cochrane Library y LILACS. La selección de estudios se realizó mediante pares de revisores, se seleccionaron estudios de pruebas diagnósticas utilizando: ELISA, ELISA indirecto, ELISA rápido, Fas2 ELISA, Arco2 comparada con técnicas de sedimentación para el diagnóstico de fascioliasis hepática en adultos. Los estudios fueron evaluados mediante la herramienta QUADAS-2. Resultados: Se identificaron siete estudios que responden a la pregunta PICO, con un total de 1317 muestras de suero humano, las pruebas reportaron alta sensibilidad y especificidad, FhTA, ELISA, número de estudios y participantes identificados, estimadores resumen con los correspondientes intervalos de confianza/credibilidad, también se puede discutir el ranking de tratamientos. Discusión: en general la especificidad de las pruebas serológicas fue ligeramente superior que la sensibilidad, existe evidencia de moderada a baja calidad que reporta su rendimiento, sin embargo, la calidad de la evidencia y la heterogeneidad entre los estudios no permite determinar su utilidad durante la fase aguda de la enfermedad. Se necesitan más estudios al respecto. Los estudios fueron clínicamente heterogéneos entre ellos.
ABSTRACT Introduction: Fascioliasis is a disease caused by the parasite Fasciola hepatica, according to its clinical picture, it can be classified into two phases: acute and chronic, currently the diagnosis is made during the chronic phase, however, early detection would allow to provide an effective treatment and timely. Objetive: This study is carried out with the objective of evaluating the diagnostic validity of serological tests for the timely diagnosis of hepatic fascioliasis. Materials and methods: a systematic search of the literature was carried out in Embase, MedLine, Cochrane Library and LILACS. Study selection was performed by pairs of reviewers, diagnostic test studies were selected using: ELISA, indirect ELISA, rapid ELISA, Fas2 ELISA, Arco2 compared to sedimentation techniques for the diagnosis of hepatic fascioliasis in adults. The studies were evaluated using the QUADAS-2 tool. Results: Seven studies were identified that answer the PICO question, with a total of 1,317 human serum samples, the tests reported high sensitivity and specificity, FhTA, ELISA, number of studies and participants identified, summary estimators with the corresponding confidence intervals / credibility, and the ranking of treatments. Discussion: In general, the specificity of the serological tests was slightly higher than the sensitivity, there is evidence of moderate to low quality that reports their performance, however, the quality of the evidence and the heterogeneity between the studies do not allow determining its usefulness during the acute phase of the disease. More studies are needed in this regard. The studies were clinically heterogeneous between them.
Assuntos
Humanos , Fasciolíase/sangue , Fasciolíase/diagnóstico , Testes SorológicosRESUMO
Abstract The lymnaeids are important in the epidemiology of Fasciola hepatica, a neglected and endemic zoonosis. The interaction between the internal defense system of Pseudosuccinea columella and F. hepatica has been little studied. In the present study the effect of infection by F. hepatica on P. columella circulating haemocytes was investigated. Changes in the average number of total circulating haemocytes have been observed at 30 minutes post-infection and 1, 7, 10, 14, 21, 28 and 50 days post-infection (dpi). Miracidia were observed head-foot and mantle at 30 minutes post-infection. Miracidia/Sporocysts in the mantle skirt 1 dpi, and fully formed sporocysts were observed in the head-foot at 7 dpi. Rediae became evident at 10 dpi and were located between the haemocoel and the muscles from 14 dpi; 50 dpi, the rediae in the digestive gland contained cercariae. The statistical analysis of the total haemocytes of P. columella infected by F. hepatica showed significant differences on the 30 minutes post-infection and 1, 14, 21, and 28 dpi in comparison to uninfected molluscs (0 dpi). Therefore, the interference observed on the internal defence system of P. columella may have direct association with the development of F. hepatica.
Resumo Os limnaeideos são importantes na epidemiologia de Fasciola hepatica, uma zoonose negligenciada e endêmica. A interação entre o sistema interno de defesa de Pseudosuccinea columella e F. hepatica tem sido pouco estudada. No presente estudo, investigou-se o efeito da infecção por F. hepatica nos hemócitos circulantes de P. columella. Alterações no número médio de hemócitos circulantes foram observadas aos 30 minutos e 1, 7, 10, 14, 21, 28 e 50 dias após a infecção (dpi). Miracídios foram observados na região cefalopodal e manto aos 30 minutos após a infecção. Miracídio/esporocistos foram observados no colar do manto ao 1 dpi, e esporocistos totalmente formados na região cefalopodal aos 7 dpi. Rédias tornam-se evidentes aos 10 dpi entre a hemocele e músculos a partir de 14 dpi; e rédias com cercárias próximas a glândula digestiva aos 50 dpi. A análise estatística dos hemócitos totais de P. columella infectados por F. hepatica demonstrou diferenças significativas nos 30 minutos pós-infecção e 1, 14, 21 e 28 dpi em comparação aos moluscos não infectados (0 dpi). Portanto, a interferência observada no sistema de defesa interna de P. columella pode ter associação direta com o desenvolvimento de F. hepatica.
Assuntos
Animais , Caramujos/parasitologia , Fasciolíase/veterinária , Hemócitos/parasitologia , Fasciolíase/sangueRESUMO
The lymnaeids are important in the epidemiology of Fasciola hepatica, a neglected and endemic zoonosis. The interaction between the internal defense system of Pseudosuccinea columella and F. hepatica has been little studied. In the present study the effect of infection by F. hepatica on P. columella circulating haemocytes was investigated. Changes in the average number of total circulating haemocytes have been observed at 30 minutes post-infection and 1, 7, 10, 14, 21, 28 and 50 days post-infection (dpi). Miracidia were observed head-foot and mantle at 30 minutes post-infection. Miracidia/Sporocysts in the mantle skirt 1 dpi, and fully formed sporocysts were observed in the head-foot at 7 dpi. Rediae became evident at 10 dpi and were located between the haemocoel and the muscles from 14 dpi; 50 dpi, the rediae in the digestive gland contained cercariae. The statistical analysis of the total haemocytes of P. columella infected by F. hepatica showed significant differences on the 30 minutes post-infection and 1, 14, 21, and 28 dpi in comparison to uninfected molluscs (0 dpi). Therefore, the interference observed on the internal defence system of P. columella may have direct association with the development of F. hepatica.
Assuntos
Fasciolíase/veterinária , Hemócitos/parasitologia , Caramujos/parasitologia , Animais , Fasciolíase/sangueRESUMO
Background: Fascioliasis is an infectious disease caused by parasites Fasciola hepatica and F. gigantica. Humans are infected by the consumption of vegetables and water contaminated with the infective form of the parasite. Materials and Methods: In this study, an IgM-ELISA with the cysteine proteinase Fas2 antigen was evaluated with sera from 76 patients infected with F. hepatica, 24 patients with other parasite infections and 84 healthy volunteers. Results: IgM-ELISA resulted in 43% positives in F. hepatica patients with positive serology to Fas2-ELISA, but no positives resulted from testing healthy volunteers and individuals infected with other parasites. The IgM-ELISA diagnostic parameters showed a sensitivity of 43.4% (95% CI 0.321-0.553), specificity of 100% (95% CI 0.957-1), and no cross-reactivity with other parasitic infection. Interference by rheumatoid factor in the IgM immunoassay was controlled by treating sera with rheumatoid factor absorbent before testing. Conclusions: Fas2 antigen is detected by circulating IgM in patients infected with F. hepatica and IgM-ELISA using Fas2 appears as a specific immunoassay to detect the acute phase of the acute phase of F. hepatica infection in humans.
Assuntos
Anticorpos Anti-Idiotípicos/sangue , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/sangue , Cisteína Endopeptidases/sangue , Fasciola hepatica/imunologia , Fasciolíase/sangue , Imunoglobulina G/sangue , Imunoglobulina M/imunologia , Animais , Área Sob a Curva , Ensaio de Imunoadsorção Enzimática , Fasciola hepatica/enzimologia , Fasciolíase/imunologia , Humanos , Fatores Imunológicos/farmacologia , Peru , Valor Preditivo dos Testes , Curva ROC , Fator Reumatoide/farmacologia , Sensibilidade e Especificidade , Estudos SoroepidemiológicosRESUMO
The enzymatic activities of NTPDase, 5'-nucleotidase and adenosine deaminase (ADA) are important in regulating the concentration of adenine nucleotides, molecules known to be involved on platelet aggregation. Fasciolosis causes coagulation disorders that have not been completely elucidated. Taking into consideration the association between the purinergic system and hemostasis, this study aimed to evaluate the enzymatic activities of NTPDase (hydrolyze ATP and ADP), 5'-nucleotidase (hydrolyze AMP) and ADA (deamination of adenosine) in platelets from cattle experimentally infected by Fasciola hepatica on days 20, 40, 60 and 80 post-infection (PI). For this study, 10 healthy Friesian steers were separated into two groups: the group A (n = 5) was used as uninfected control, and the group B was composed of steers experimentally infected by F. hepatica (n = 5). The number of platelets did not differ between groups in the periods evaluated. Reduction of NTPDase (p < 0.05) hydrolysing ATP (days 20, 40 and 60 PI), and ADP (days 40, 60 and 80 PI), and on 5'-nucleotidase hydrolyzing AMP (days 40 and 60 PI) was observed. A reduction (p < 0.05) in ADA activity on day 20 PI, as well as an increase (p < 0.05) in ADA activity on days 40 and 60 PI was observed when compared to the control. Based on these results, we can conclude that ATP, ADP and AMP hydrolysis and adenosine deamination were altered in platelets of cattle infected by F. hepatica. Considering the importance of the purinergic system in hemostasis, it is believed that those changes may contribute to the coagulation impairment observed in acute fasciolosis described in the literature.
Assuntos
Adenosina Desaminase/sangue , Plaquetas/enzimologia , Doenças dos Bovinos/parasitologia , Fasciolíase/veterinária , Nucleotidases/sangue , Animais , Bovinos , Doenças dos Bovinos/sangue , Doenças dos Bovinos/enzimologia , Fasciola hepatica/fisiologia , Fasciolíase/sangue , Fasciolíase/enzimologia , Fezes/parasitologia , Fígado/parasitologia , Fígado/patologia , Masculino , Contagem de Ovos de Parasitas/veterinária , Contagem de Plaquetas/veterináriaRESUMO
The objective of present study was to determine the prevalence of natural caprine fasciolosis in the Mixteca region of Mexico using coproantigen and serum IgG1 ELISA tests for comparative purposes. A total of 1070 serum and faecal samples were analyzed for IgG1 antibodies and coproantigens, using ELISA with E/S products as antigen and a monoclonal antibody-based sandwich ELISA. Prevalence of 73.46% was found using the serological ELISA and a percentage of 77.20 was found for coproantigen ELISA. The diagnostic sensitivity and specificity for serum ELISA were 86.7% and 96.4%, and for the coproantigen ELISA they were 93.1% and 97.8%, respectively. The seropositive samples were further categorized as low, medium, or high positivity. Results show a great proportion of low and medium positive goats when the serum ELISA test was used. Correlation coefficients between coproantigens and seropositivity were statistically significant (P < 0.01) for low seropositivity (r = 0.93) and medium seropositivity (r = 0.84). The accuracy of faecal antigen ELISA was higher compared to indirect ELISA serological test. Two ELISAs were shown to be useful for demonstrating the current status of F. hepatica infection in the endemic areas and can be employed in studies on epidemiology as well as anthelmintics treatment for preventing economic loss and the risk of transmission to humans.
Assuntos
Ensaio de Imunoadsorção Enzimática/veterinária , Fasciolíase/veterinária , Doenças das Cabras/sangue , Doenças das Cabras/diagnóstico , Imunoglobulina G/sangue , Animais , Antígenos/imunologia , Biomarcadores/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Fasciolíase/sangue , Fasciolíase/diagnóstico , Feminino , Doenças das Cabras/imunologia , Cabras/sangue , Cabras/imunologia , Imunoglobulina G/imunologia , Masculino , México , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
OBJECTIVES: To evaluate the performance of the enzyme-linked immunoelectrotransfer blot assay (EITB, Western blot) using excretory/secretory antigens from adult forms of Fasciola hepatica (Fh E/S Ag) for the diagnosis of human fasciolosis. MATERIALS AND METHODS: Antigens were obtained after 18 hours of incubation in culture medium Minimum Essential Eagle, prepared at a protein concentration of 0.15 ug/uL and run against a pool of sera of patients with proven fasciolosis (confirmed by the finding of parasite eggs in the stool microscopy). Antigens of 10, 12, 17, 23, 27, 30, 36, 43, 66 and 136 kDa were detected and used to develop the Western blot technique. The sensitivity was evaluated using sera from 67 fasciolosis patients, and the specificity using sera from 57 patients with other parasitic diseases, and 10 from healthy individuals. RESULTS: Out of the 67 sera, 64 reacted with the 23 kDa band and 61 with the one of 17 kDa. These two bands were not detected in sera from patients with other parasitic diseases or in those from healthy volunteers and thus could be considered specific and diagnostic. CONCLUSIONS: The sensitivity of the test, using the bands of 17 and 23 kDa, was 95.5% for positive reactions to at least one of these two bands, being its specificity 100% with a positive predictive value of 100% and negative predictive value of 95.71%.
Assuntos
Antígenos de Helmintos/análise , Western Blotting/normas , Fasciola hepatica/imunologia , Fasciolíase/sangue , Fasciolíase/diagnóstico , Adolescente , Adulto , Animais , Criança , Fezes/parasitologia , Humanos , Pessoa de Meia-Idade , Adulto JovemRESUMO
Objetivos. Evaluar la eficacia de la técnica de electroinmunotransferencia (EITB) o Western blot utilizando antígenos de excreción-secreción de las formas adultas de Fasciola hepatica (Fh E/S Ag) para el diagnóstico de la fasciolosis humana. Materiales y métodos. Los antígenos fueron obtenidos a las 18 horas de incubación en medio Minimum Essential Eagle y preparados a la concentración proteica de 0,15 ug/uL; los cuales, al ser enfrentados con un pool de sueros de pacientes con fasciolosis confirmada por el hallazgo de huevos del parásito en las heces, se detectaron los antígenos de 10, 12, 17, 23, 27, 30, 36, 43, 66 y 136 KDa, con los cuales se desarrolló la técnica de Western blot. La sensibilidad se evaluó empleando sueros de 67 pacientes con fasciolosis, y la especificidad con sueros de 57 pacientes con otras parasitosis y diez sueros de personas no parasitadas. Resultados. De los 67 sueros, 64 reaccionaron con la banda de 23 KDa y 61 con la banda de 17KDa. Estas dos bandas no fueron detectadas por ninguno de los sueros de pacientes con otras parasitosis, ni de personas no parasitadas, siendo por ello consideradas como específicas y diagnósticas. Conclusiones. La sensibilidad de la prueba, utilizando las bandas de 17 y 23 KDa, fue de 95,5 por ciento cuando se presenta reacción positiva en una o en las dos bandas, siendo la especificidad para estos dos antígenos de 100 por ciento con un valor predictivo positivo de 100 por ciento y un valor predictivo negativo de 95,71 por ciento.
Objectives. To evaluate the performance of the enzyme-linked immunoelectrotransfer blot assay (EITB, Western blot) using excretory/secretory antigens from adult forms of Fasciola hepatica (Fh E/S Ag) for the diagnosis of human fasciolosis. Materials and methods. Antigens were obtained after 18 hours of incubation in culture medium Minimum Essential Eagle, prepared at a protein concentration of 0.15 ug/uL and run against a pool of sera of patients with proven fasciolosis (confirmed by the finding of parasite eggs in the stool microscopy). Antigens of 10, 12, 17, 23, 27, 30, 36, 43, 66 and 136 kDa were detected and used to develop the Western blot technique. The sensitivity was evaluated using sera from 67 fasciolosis patients, and the specificity using sera from 57 patients with other parasitic diseases, and 10 from healthy individuals. Results. Out of the 67 sera, 64 reacted with the 23 kDa band and 61 with the one of 17 kDa. These two bands were not detected in sera from patients with other parasitic diseases or in those from healthy volunteers and thus could be considered specific and diagnostic. Conclusions. The sensitivity of the test, using the bands of 17 and 23 kDa, was 95.5 percent for positive reactions to at least one of these two bands, being its specificity 100 percent with a positive predictive value of 100 percent and negative predictive value of 95.71 percent.
Assuntos
Adolescente , Adulto , Animais , Criança , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Antígenos de Helmintos/análise , Western Blotting/normas , Fasciola hepatica/imunologia , Fasciolíase/sangue , Fasciolíase/diagnóstico , Fezes/parasitologiaRESUMO
Se obtuvo el antígeno metabólico (antígeno excreción - secreción) de Fasciola hepatica de ovinos infectados de Cajamarca, con una concentración proteica de 1 005 μg/μL, compuesta principalmente por proteνnas de peso molecular entre 1,2 y 170 KDa. Se detectaron bandas de 170; 150; 31; 24; 18-14 y 10 kDa. Con este antνgeno se desarrollσ una prueba de ELISA y se determinσ su punto de corte en 0,140. Se evaluσ 33 sueros de pacientes con fasciolosis confirmada por visualización de huevos en heces, 177 sueros de pacientes sin fasciolosis provenientes de áreas endémicas de Cajamarca y 88 sueros de pacientes con otras infecciones parasitarias y bacterianas. Se encontró una sensibilidad de 97,0 por ciento, especificidad de 96,6 por ciento, valor predictivo positivo de 78,1 por ciento y valor predictivo negativo de 99,6 por ciento. Se encontró reacción cruzada en 9/88 sueros evaluados. Se recomienda la implementación y uso de esta prueba para el diagnóstico de fasciolosis.
Metabolic (excretion/secretion) antigen was obtained from sheep infected with Fasciola hepatica, with a 1005 μg/μL of protein concentration, composed principally by proteins of molecular weight between 1.2 and 170 KDa. Bands of 170, 150, 31, 24, 18-14 and 10 kDa were detected. With this antigen an ELISA test was developed and the cut off was determined in 0.140. We evaluated 33 serums of patient with fascioliasis confirmed by visualization of eggs in feces, 177 serums of persons without fascioliasis from endemic rural areas of Cajamarca and 88 serums of patients with others parasitic and bacterial infections. We found a 97.0 percent of sensitivity, 96.6 specificity, 78.1 percent predictive positive value, 99.6 percent predictive negative value. In 9/88 serums was found cross reactions. We recommended the implementation and use of this test for the fascioliasis diagnosis.
Assuntos
Animais , Humanos , Antígenos de Helmintos/sangue , Ensaio de Imunoadsorção Enzimática , Fasciola hepatica/imunologia , Fasciolíase/sangue , Fasciolíase/diagnóstico , Peru , Sensibilidade e EspecificidadeRESUMO
Metabolic (excretion/secretion) antigen was obtained from sheep infected with Fasciola hepatica, with a 1005 µg/µL of protein concentration, composed principally by proteins of molecular weight between 1.2 and 170 KDa. Bands of 170, 150, 31, 24, 18-14 and 10 kDa were detected. With this antigen an ELISA test was developed and the cut off was determined in 0.140. We evaluated 33 serums of patient with fascioliasis confirmed by visualization of eggs in feces, 177 serums of persons without fascioliasis from endemic rural areas of Cajamarca and 88 serums of patients with others parasitic and bacterial infections. We found a 97.0% of sensitivity, 96.6 specificity, 78.1% predictive positive value, 99.6 % predictive negative value. In 9/88 serums was found cross reactions. We recommended the implementation and use of this test for the fascioliasis diagnosis.
Assuntos
Antígenos de Helmintos/sangue , Ensaio de Imunoadsorção Enzimática , Fasciola hepatica/imunologia , Fasciolíase/sangue , Fasciolíase/diagnóstico , Animais , Humanos , Peru , Sensibilidade e EspecificidadeRESUMO
An epidemiological focal study was performed in Loncopué, Neuquén, Argentina, in November 2002 to detect the origin of the infection in a human case of fascioliasis confirmed by an indirect-ELISA test, six months before the study. Thirty five individual fecal samples were taken from domestic livestock, and watercress plants and snails were collected from the irrigation ditches connected to a main canal in the surroundings of the patient's house. A new blood sample was taken from the already recovered patient. The patient was still seropositive to Fasciola hepatica antigens. No metacercariae were found in the 222 watercress leaves checked. All the snails collected (n=130) were identified as Lymnaea viatrix and two out of 101 (2%) were infected with F. hepatica larvae. Coprological analysis showed F. hepatica eggs in 100% of goats (10/10), 82% of sheep (9/11) and 86% of bovines (6/7). The number of eggs per gram shed by positive goats (median=20.7, Q1=6.2, Q3=34.5) and sheep (4, 1.8, 13) was significantly higher than in cows (0.3, 0.3, 1.7) (p < 0.01). Local veterinary control programs were apparently not effective in this case. Anthelmintics used and treatment schedule should be revised and small herds raised at households should also be included and treated.
Assuntos
Doenças dos Bovinos/parasitologia , Fasciola hepatica/isolamento & purificação , Fasciolíase/etiologia , Adulto , Animais , Bovinos , Ensaio de Imunoadsorção Enzimática , Fasciolíase/sangue , Fasciolíase/veterinária , Fezes/parasitologia , Feminino , Parasitologia de Alimentos , Cabras , Humanos , Lymnaea/parasitologia , Nasturtium/parasitologia , Contagem de Ovos de Parasitas , Ovinos , Estatísticas não ParamétricasRESUMO
El objetivo del trabajo fue realizar un estudio de foco relacionado con un caso de fasciolosis humana ocurrido en abril de 2002 en Loncopué, Neuquén, Argentina. La confirmación diagnóstica se efectuó en mayo de 2002 por el test de ELISA. En noviembre de 2002 se realizó un muestreo en el área rural donde se ubicaba la vivienda de la paciente, ya restablecida, y se le tomó una nueva muestra de sangre. El suero de la paciente continuó reactivo para antígenos de Fasciola hepatica. Se muestrearon plantas de berro para detectar metacercarias. Se recolectaron caracoles en cuatro canales de riego conectados a un canal principal. Los caracoles fueron trasladados vivos para su identificación, medición y examen de infección. Se recolectaron 35 muestras fecales de ganado de cría. No se observaron metacercarias en las hojas de berro examinadas (n=222). Se recolectaron 130 caracoles identificados como Lymnaea viatrix y 2 de 101 ejemplares (2%) estaban infectados con larvas de F. hepatica. Las prevalencias en el ganado adulto fueron: 100% (10/10) para caprinos, 82% (9/11) para ovinos y 86% (6/7) para bovinos. El número de huevos eliminados por las cabras (mediana = 20.7; Q1=6.2; Q3=34.5) y ovejas (4, 18, 13) infectadas, resultó mayor que el eliminado por vacas (0.3; 0.3; 1.7) (p<0.01). La práctica de control local no tuvo efecto aparente en este caso, por lo que deberían revisarse los calendarios de tratamiento y los antiparasitarios utilizados. Los resultados muestran que el ganado criado a pequeña escala por los pobladores debe incluirse en los programas de control. Se discute la posible importancia de la fasciolosis humana en Argentina.
Assuntos
Adulto , Bovinos , Animais , Humanos , Feminino , Doenças dos Bovinos/parasitologia , Fasciola hepatica/isolamento & purificação , Fasciolíase/veterinária , Ensaio de Imunoadsorção Enzimática , Parasitologia de Alimentos , Fasciolíase/sangue , Fezes/parasitologia , Cabras , Lymnaea/parasitologia , Nasturtium/parasitologia , Contagem de Ovos de Parasitas , Ovinos , Estatísticas não ParamétricasRESUMO
El objetivo del trabajo fue realizar un estudio de foco relacionado con un caso de fasciolosis humana ocurrido en abril de 2002 en Loncopué, Neuquén, Argentina. La confirmación diagnóstica se efectuó en mayo de 2002 por el test de ELISA. En noviembre de 2002 se realizó un muestreo en el área rural donde se ubicaba la vivienda de la paciente, ya restablecida, y se le tomó una nueva muestra de sangre. El suero de la paciente continuó reactivo para antígenos de Fasciola hepatica. Se muestrearon plantas de berro para detectar metacercarias. Se recolectaron caracoles en cuatro canales de riego conectados a un canal principal. Los caracoles fueron trasladados vivos para su identificación, medición y examen de infección. Se recolectaron 35 muestras fecales de ganado de cría. No se observaron metacercarias en las hojas de berro examinadas (n=222). Se recolectaron 130 caracoles identificados como Lymnaea viatrix y 2 de 101 ejemplares (2%) estaban infectados con larvas de F. hepatica. Las prevalencias en el ganado adulto fueron: 100% (10/10) para caprinos, 82% (9/11) para ovinos y 86% (6/7) para bovinos. El número de huevos eliminados por las cabras (mediana = 20.7; Q1=6.2; Q3=34.5) y ovejas (4, 18, 13) infectadas, resultó mayor que el eliminado por vacas (0.3; 0.3; 1.7) (p<0.01). La práctica de control local no tuvo efecto aparente en este caso, por lo que deberían revisarse los calendarios de tratamiento y los antiparasitarios utilizados. Los resultados muestran que el ganado criado a pequeña escala por los pobladores debe incluirse en los programas de control. Se discute la posible importancia de la fasciolosis humana en Argentina. (AU)
Assuntos
Adulto , Bovinos , Animais , Humanos , Feminino , Fasciolíase/veterinária , Doenças dos Bovinos/parasitologia , Fasciola hepatica/isolamento & purificação , Fasciolíase/sangue , Nasturtium/parasitologia , Parasitologia de Alimentos , Lymnaea/parasitologia , Fezes/parasitologia , Contagem de Ovos de Parasitas , Ensaio de Imunoadsorção Enzimática , Estatísticas não Paramétricas , Cabras , OvinosRESUMO
This is a retrospective and descriptive review of 277 patients suffering fasciolasis. These patients were seen in Hospital Nacional Cayetano Heredia between 1970-2002; 240 (86.6%) developed the chronic phase and 37 (13.3%) the acute form. Group aged 20-29 years were the most affected (24%). The main places infected were the interandean valleys of Lima, Ancash and Junin. The 277 patients the main symptoms were: abdominal pain in 236, nausea/vomiting in 106 and diarrhea in 100. Hepatomegaly in 56, pallor in 44 and fever in 23 were the main physical findings. 47% (80/169) suffered eosinophilia, 46% (38/82) had an increase of alkaline phosphatase and 31% (52/169) anemia.
Assuntos
Fasciola hepatica/isolamento & purificação , Fasciolíase/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Animais , Análise Química do Sangue , Criança , Pré-Escolar , Fasciolíase/sangue , Fasciolíase/patologia , Fezes/parasitologia , Feminino , Testes Hematológicos , Humanos , Masculino , Pessoa de Meia-Idade , Contagem de Ovos de Parasitas , Peru/epidemiologia , Estudos RetrospectivosRESUMO
The aim of this study was to evaluate the kinetics of the cytokines interferon-gamma, interleukin-2, interleukin-10 and interleukin-4 produced by spleen mononuclear cells stimulated by Con A during an experimental infection in rats with Fasciola hepatica. The proliferative response to Con A of Spm cells from rats infected with F. hepatica was significantly decreased on day 7 post-infection (P<0.006) and simultaneously an increase of interferon-gamma, interleukin-10 and interleukin-4 production along with a decrease of interleukin-2 by spleen mononuclear cells were observed. Interleukin-4 and interleukin-10 were involved in ablating cellular proliferation in vitro, as the addition of neutralising antibodies to either cytokine reversed the proliferative block. The addition of exogenous recombinant interleukin-2 also restored the proliferative response by spleen mononuclear cells obtained 7 days after infection from infected rats. At the same time, we found an increase in interleukin-10 production by peritoneal cells (in close contact with the flukes) and decreased nitric oxide levels. In addition, histological studies on the liver on day 7 after infection showed the presence of parasite inside migratory tunnels in the parenchyma, and polymorphonuclear leukocytes, predominantly eosinophils, around the parasite. The transient suppression in proliferative response mediated by cytokines interleukin-4 and interleukin-10 in the spleen, and diminution of nitric oxide production in the peritoneum could be mechanisms to evade the protective immune response during the first stages of liver penetration by the parasite.
Assuntos
Citocinas/biossíntese , Fasciola hepatica/imunologia , Fasciolíase/imunologia , Animais , Concanavalina A/imunologia , Citocinas/sangue , Fasciolíase/sangue , Feminino , Histocitoquímica , Interferon gama/biossíntese , Interferon gama/sangue , Interleucinas/biossíntese , Interleucinas/sangue , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Fígado/parasitologia , Fígado/patologia , Macrófagos Peritoneais/imunologia , Óxido Nítrico/análise , Óxido Nítrico/biossíntese , Ratos , Ratos Wistar , Baço/citologia , Baço/imunologia , Baço/metabolismoRESUMO
Enzyme-linked immunosorbent assay (ELISA) and micro-ELISA were evaluated for their ability to detect anti-Fasciola hepatica antibodies in humans by using excretory-secretory antigen. The sensitivity of each method was 100%, but the specificity was 100% for ELISA and 97% for micro-ELISA. The micro-ELISA could be used as a screening assay and ELISA could be used as a confirmatory method for the serodiagnosis of human fascioliasis.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Fasciolíase/diagnóstico , Animais , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/imunologia , Bovinos , Fasciolíase/sangue , Fasciolíase/imunologia , Humanos , Testes Imunológicos/métodosRESUMO
Los objetivos del presente estudio fueron: Determinar la sensibilidad y especificidad de DIG-ELISA y establecer el valor de corte para realizar diagnóstico de fasciolosis bovina; determinar la capacidad diagnóstica de la prueba en bovinos infectados en forma experimental; valorar la seroprevalencia bimensual y durante un año de los títulos IgG anti-Fasciola hepatica en suero de bovinos infectados en forma natural; y comparar la variación con la presencia de huevos del trematodo identificados en el análisis coproparasitoscópico. Con el propósito de estandarizar la prueba, además de las correspondientes titulaciones de sueros, antígeno y conjugado, se utilizaron 80 sueros de bovinos, de estos últimos 40 pertenecían a animales provenientes del estado de Sonora (zona libre de fasciolosis) y 40 del estado de Hidalgo (zona enzoótica de fasciolosis). La prueba mostró una sensibilidad de 97.5 por ciento y una especificidad de 80.0 por ciento, obteniendo un valor de corte de 7 mm. En otro estudio, se infectaron 15 bovinos jóvenes de sexo indistinto, cada uno con 600 metacercarias del Fasciola hepatica. Los animales fueron sangrados semanalmente y durante 14 ocasiones para obtención de suero. La prueba fue capaz de detectar anticuerpos anti-F. Hepatica desde la segunda semana posinfección, logrando sus mayores incrementos expresados en diámetro de la zona de reacción (DZR) entre la séptima y décima semanas. En otro experimentos se utilizaron 85 bovinos criollos, hembras, de un rancho ubicado en un área enzoótica de fasciolosis en Nautla, Veracruz, en donde se colectó sangre para obtención de suero, así como heces de cada uno de los bovinos. Los animales fueron muestreados bimensualmente y durante un año para correr los cueros con DIG-ELISA y las heces para realizar análisis por sedimentación. Los porcentajes de seroprevalencia fueron del 58 por ciento, 73 por ciento, 80 por ciento, 100 por ciento, 96 por ciento y 79 por ciento para los meses de marzo, mayo, julio, septiembre, enero y marzo, respectivamente, mostrando porcentajes de positividad similares a aquellos obtenidos en el análisis coproparasitoscópico (P>0.01). Se concluye que DIG-ELISA mostró alta sensibilidad y fácil montaje para utilizarse bajo condiciones de campo en el diagnóstico temprano de fasciolisis bovina, generando resultados reproducibles que pueden ser de gran utilidad para estudios seroepidemiológicos